AUTHOR=Shukla Alka , Gangwar Mayank , Sharma Gaurav , Prakash Pradyot , Nath Gopal 

TITLE=Vitality of Proteinase K in rRTPCR Detection of SARS-CoV2 Bypassing RNA Extraction

JOURNAL=Frontiers in Cellular and Infection Microbiology

VOLUME=Volume 11 - 2021

YEAR=2021

URL=https://www.frontiersin.org/journals/cellular-and-infection-microbiology/articles/10.3389/fcimb.2021.717068

DOI=10.3389/fcimb.2021.717068

ISSN=2235-2988

ABSTRACT=This study aimed to detect SARS-COV2 viral component directly from inoculated VTM without RNA extraction. Inoculated VTMs of already tested 50 positives and 50 negatives were grouped into three groups. Group I was treated with Proteinase K (PK) followed by 3-step-heat treatment at different temperatures (25˚C, 60˚C and 98˚C) and stored at 4˚C. Group II was directly subjected to 3-step-heat treatment without PK exposure and stored at 4˚C. And group III was set-up as standard group and it was processed using Qiagen’s column based QIAamp Nucleic Acid kit and the obtained nucleic acids were stored at 4˚C. These stored samples were used as a template to execute real-time polymerase chain reaction, and results were noted down. Group I demonstrated 96% and 88% sensitivity for N and ORF1ab genes respectively, whereas group II demonstrated 78% and 60% when compared to results of standard group III. Overall group I showed better results than group II when compared to group III. Thus, in situations where gold-standard reagents are not available, PK exposure and heat treatment can be employed to carry out molecular detection of SARS-CoV2 viral component.